Medicinal plants have been used as an alternative in malaria chemotherapy. This study aimed to determine the in vitro inhibitory potentials of the extract and fractions of Brenania brieyi on Lactate Dehydrogenase (LdH), Phospholipase A2 (PLA2), and α-Amylase enzyme extracted from Plasmodium falciparum. Cold maceration and Liquid-Liquid partitioning were used to obtain the extract and fractions respectively, using different solvent of varying polarity. Phytochemical analysis was done using Gas Chromatography-Flame Ionization detector (GCFID) method to identify and quantify different phytoconstituents. In vitro enzyme assay (α-Amylase, LdH and PLA2) were performed using standard procedure. The extract contained Alkaloids such as Spartein (5.05 µg/ml), Dihydrocytisine (23.12 µg/ml), Aphyllidine (5.25 µg/ml), Ribalinidine (2.65 µg/ml), Epihedrine (1.91 µg/ml), Saponins such as sapogenin (5.70 µg/ml), Tannins (16.25 µg/ml), Flavonoids such as Kaempferol (4.59 µg/ml), Catechin (5.50 µg/ml), Proanthocyanidin (9.81 µg/ml), Anthocyanin (3.22 µg/ml), Narigenin (3.62 µg/ml), Flavonones (8.73 µg/ml), Steroids (8.58 µg/ml), Polyphenols such as Reservatrol (20.68 µg/ml) and Glycosides such as cardiac glycoside (4.42 µg/ml), Cyanogenic glycosides (17.72 µg/ml) while Terpenoids are absent. The percentage inhibition of the enzyme (PLA2, LdH and α-amylase) at 100 mg/ml of extract showed the highest inhibition (37 %, 0 % and 38.33 %) when compared to controls, while ethyl acetate fraction has the highest inhibition (98 %, 34 % and 92 %) at P ˂ 0.05 at 100 mg/kg respectively. This study justifies the use of ethnomedicinal remedies (B. brieyi) in treating malaria in infected individuals. The study also suggests that the extract and fractions may act through the inhibition of plasmodial enzymes in infected persons.